Fluorescent bacteria were isolated from sporocarps that browned into various mushrooms during survey at places of the production in Korea. We examined the pathogenicity, biodiversity, and genetic characteristics of the 19 strains identified as Pseudomonas tolaasii by sequence analysis of 16S rRNA and White Line Assay. The results emphasize the importance of rpoB gene system, fatty acid profiles, specific and sensitive PCR assays, and lipopeptide detection for the identification of P. tolaasii. As a result of these various analyses, 17 strains (CHM03~CHM19) were identified as P. tolaasii. The phylogenetic analysis based on the 16S rRNA gene showed that all strains were clustered closest to P. tolaasii lineage, two strains (CHM01, CHM02) were not identified as P. tolaasii and have completely different genetic characteristics as a result of fatty acids profile, specific and sensitive PCR, lipopetide detection, rpoB sequence and REP-PCR analysis. Pathogenicity tests showed 17 strains produce severe brown discolouration symptoms to button mushrooms and watersoaking of sporophore tissue within three days after inoculation. But two strains did not produce discolouration symptoms. Therefore, these two strains will be further investigated for correct species identification by different biological and molecular characteristics.

팽이버섯 재배를 하고 있는 농가의 배지조성을 조사한 결과 농가마다 사용하는 배지재료의 종류와 혼합비율도 차이가 있었다. 모든 농가가 콘코브, 미강, 비트펄프를 주 재료로 사용하였고, 배지의 적정 pH 조절을 위하여 탄산 칼슘과 폐화석을 1.0~3.8%을 사용하였으며, 건비지, 밀기울, 면실피, 파옥쇄 등은 농가마다 사용하는 정도가 달랐다. 농가별 혼합배지의 이화학성을 분석한 결과 질소함량은 1.28~1.52%였고, 탄소함량은 45.7~48.5%, C/N율은 30.7~37.9였다. 무기성분 함량은 혼합배지에 들어가는 무기물의 종류와 양에 따라 농가마다 큰 차이를 보였다. 균사 생장량은 배양 34일후 농가 2에서 6.5 cm로 가장 빨랐고, 농가 4에서는 5.7 cm였으며, 농가 1, 농가 3, 농가 5에서는 균사 생장량이 건의 비슷하였다. 균사 밀도는 처리 간에 뚜렷한 차이가 없이 높은 밀도를 보였다. 농가별 배지조성에 따른 수량성을 조사한 결과 농가2에서 173.4 g/ bottle로 가장 높은 수량을 보였으며, 농가 1에서는 168.4 g/bottle이였고, 농가 3과 4는 거의 비슷한 수량을 보였다. 그러나 농가 5는 145.4 g/bottle로 가장 낮은 수량을 보였다. 이와 같이 농가별 배지조성에 따라 팽이버섯의 수량이 차이를 보였으며, 이러한 결과를 참고하여 팽이버섯 재배농가에 범용적으로 사용할 수 있는 배지조성의 선발이 가능할 것으로 판단된다.

The button mushroom, Agaricus bisporus, is one of the major economical crops cultivated in Korea. This mushroom showed the 5th production to 11,493 M/T in 2014. This study examined changes in decomposition-related enzyme activity secreted by the microorganisms isolated from button mushroom composts. Among the isolated microorganisms, bacterial strains CY-36 were the most dominant and had the highest enzyme activities. 16S rDNA sequencing analysis showed 99% similarity of CY-36 to Bacillus amyloliquefaciens. To define the optimum conditions for the mass production of CY-36, we have investigated optimum culture conditions and effects of various nutrient source on the bacterial growth. The optimum initial pH and temperature were determined as pH 5.0~8.0 and 30°C, respectively. The optimal culture medium for the growth was determined as follows: 2.0% dextrin, 1.0% yest extract, 0.5% NaNO3, 3 mM MgSO4, and 0.6% valine. Enzymatic activities of CMCase and PGase secreted by CY-36 were the most prominent and CMCase activity were optimal at 40°C and pH 7.0, and PGase activity were 50°C and pH 6.0. The activity of CMCase was rapidly decreased at more than 4°C and pH 5.0, and the activity of PGase was rapidly decreased at more than 30°C but didn’t affect as pH change. Accordingly, Bacillus amyloliquefaciens CY-36 was potentially useful on the fermentation stimulation and softening of rice straw in the process of fermentation for mushroom cultivation.

This study examined changes in decomposition-related enzyme activity secreted by the microorganisms isolated from button mushroom composts. Among the isolated microorganisms, bacterial strains CY-24 were the most dominant and had the highest enzyme activities. 16S rDNA sequencing analysis showed 97% similarity of CY-24 to Bacillus licheniformis. To define the optimum conditions for the mass production of CY-24, we have investigated optimum culture conditions and effects of various nutrient source on the bacterial growth. The optimum initial pH and temperature were determined as pH 6.0 and 30°C, respectively. The optimal culture medium for the growth was determined as follows: 0.3% mannose, 1.5% yest extract, 0.8% (NH4)2HPO4, 2 mM MnSO4, and 0.5% asparatic acid. Enzymatic activities of CMCase and PGase secreted by CY-24 were the most prominent and CMCase activity were optimal at 50~60°C and pH 6.0, and PGase activity were 60°C and pH 6.0. The activity of CMCase was inhibited by metal ions including Ca+ and Cu+, while the activity of PGase didn’t affect. The activity of CMCase was rapidly decreased at more than 30°C and pH 7.0, and the activity of PGase didn’t affect as temperature change but was rapidly decreased at more than pH 5.0. Accordingly, This strain was potentially useful on the fermentation stimulation and softening of rice straw in the process of fermentation for mushroom cultivation.